ER,PgR, HER-2, Ki-67, topoisomerase IIα, and nm23-H1 proteins expression as predictors of pathological complete response to neoadjuvant chemotherapy for locally advanced breast cancer

The purpose of this study was to evaluate the importance of biological markers to predict pathologic complete response (pCR) to neoadjuvant chemotherapy (NCT) in patients with locally advanced breast cancer (LABC) One hundred and twelve consecutive patients with clinical stage III LABC who had received NCT with docetaxel and epirubicin from March 2006 to March 2009 were included in this retrospective study. The pre-NCT treatment expression levels of Ki-67 proliferation index, estrogen receptor (ER), progesterone receptor (PgR), epidermal growth factor receptor 2 (HER-2), Topoisomerase II alpha (Topo-II), and nm23-H1 were detected by immunohistochemistry (IHC). A total of 361 cycles were administered with the median number of three cycles per patient (range, 2–6). The pCR rate was 9.8% (95% CI, 4.3–15.3%). In univariate analysis, poor tumor differentiation, both negative of ER/PgR, negative Topo-II, and positive nm23-H1 were found to be significantly predictive of a pCR. ER/PgR status and nm23-H1 were significant for pCR on multivariate analysis (P = 0.006 and 0.025, respectively). ER/PgR status and nm23-H1 are independent predictive factors of pCR to neoadjuvant docetaxel plus epirubicin combination chemotherapy in patients with LABC.     

CK5/6, EGFR, Ki-67, cyclin D1, and nm23-H1 protein expressions as predictors of pathological complete response to neoadjuvant chemotherapy in triple-negative breast cancer patients

The purpose of this study was to evaluate the importance of biological markers to predict pathologic complete response (pCR) to neoadjuvant chemotherapy (NCT) in patients with locally advanced triple-negative breast cancers (TNBCs). Forty-one patients (18.6%) among 220 breast cancer patients were identified as TNBCs from March 2006 to 2009 were included in this prospective study. The pre-NCT treatment expression levels of Ki-67 proliferation index, estrogen receptor (ER), progesterone receptor (PgR), epidermal growth factor receptor 2 (HER-2), CK5/6, epidermal growth factor receptor (EGFR), cyclin D1, and nm23-H1 were detected by immunohistochemistry (IHC). A total of 180 cycles were administered with the median number of four cycles per patient (range, 4-6). The pCR rate was 34.1% (95% CI, 19.6-48.6%). In univariate analysis, early T stage, clinical response after 2 cycles, negative basal-like, negative EGFR, high Ki-67 proliferation index, and positive nm23-H1 were found to be significantly predictive of a pCR (P = 0.010, 0.040, 0.007, 0.001, 0.019, and 0.010, respectively). Basal-like status and nm23-H1 status were significant for pCR on multivariate analysis (P = 0.004 and 0.031, respectively). Basal-like status and nm23-H1 are independent predictive factors of pCR to neoadjuvant docetaxel plus epirubicin combination chemotherapy in patients with TNBCs.     

CK5/6, EGFR,Ki-67, cyclin D1, and nm23-H1 protein expressions as predictors of pathological complete response to neoadjuvant chemotherapy in triple-negative breast cancer patients

The purpose of this study was to evaluate the importance of biological markers to predict pathologic complete response (pCR) to neoadjuvant chemotherapy (NCT) in patients with locally advanced triple-negative breast cancers (TNBCs). Forty-one patients (18.6%) among 220 breast cancer patients were identified as TNBCs from March 2006 to 2009 were included in this prospective study. The pre-NCT treatment expression levels of Ki-67 proliferation index, estrogen receptor (ER), progesterone receptor (PgR), epidermal growth factor receptor 2 (HER-2), CK5/6, epidermal growth factor receptor (EGFR), cyclin D1, and nm23-H1 were detected by immunohistochemistry (IHC). A total of 180 cycles were administered with the median number of four cycles per patient (range, 4–6). The pCR rate was 34.1% (95% CI, 19.6–48.6%). In univariate analysis, early T stage, clinical response after 2 cycles, negative basal-like, negative EGFR, high Ki-67 proliferation index, and positive nm23-H1 were found to be significantly predictive of a pCR (P = 0.010, 0.040, 0.007, 0.001, 0.019, and 0.010, respectively). Basal-like status and nm23-H1 status were significant for pCR on multivariate analysis (P = 0.004 and 0.031, respectively). Basal-like status and nm23-H1 are independent predictive factors of pCR to neoadjuvant docetaxel plus epirubicin combination chemotherapy in patients with TNBCs.     

大肠癌中nm23-H1的表达与淋巴结转移的关系

 目的　探讨大肠癌中nm23-H1的表达与淋巴转移的关系。方法　应用免疫组化方法研究96例大肠癌中nm23-H1蛋白的表达。结果　nm23-H1蛋白低表达与淋巴结或远处转移显著相关(P<0.05);nm23-H1蛋白低表达预测大肠癌转移的灵敏性为88.4%,特异性为79.3%。结论　检测nm23-H1蛋白可以预测大肠癌淋巴结或远处转移,从而可能成为临床治疗的判断依据。     

nm23-H1 expression in salivary adenoid cystic carcinoma in relation to metastasis and survival

The expression of nm23-H1, product of putative metastasis suppressor gene, was evaluated immunohistochemically in 31 cases of adenoid cystic carcinoma (ACC) of salivary glands and correlated with their clinicopathologic features. All benign salivary gland tumors of various types, which were used as a non-metastatic control, showed obvious nm23-H1 expression. The immunoreactivity of tumor cells was stronger than that of normal salivary gland components, although the distribution patterns of positive cells considerably varied between tumor types. In ACC, 16 cases (52%) showed the reduction of nm23-H1 immunoreactivity either in positive cell frequency or staining intensity. These cases were referred to as negative cases. The incidence of negative cases was 67% (10/15) and 38% (6/16) of the cases with and without metastasis, respectively. Furthermore, metastatic tumors showed decreased immunoreactivity of this protein compared with their primary tumors. The prognosis of patients with a nm23 negative tumor was generally poorer than that with a positive tumor. These results may suggest that the reduction of nm23-H1 protein has an implication for metastasis of ACC.     

Evaluation of ER,PgR, HER-2, Ki-67, cyclin D1, and nm23-H1 as predictors of pathological complete response to neoadjuvant chemotherapy for locally advanced breast cancer

The purpose of this study was to evaluate the importance of biological markers to predict pathologic complete response (pCR) to neoadjuvant docetaxel plus epirubicin combination chemotherapy in patients with locally advanced breast cancer (LABC). Two hundred and twenty consecutive patients with LABC who had received neoadjuvant chemotherapy (NCT) with docetaxel and epirubicin from March 2006 to March 2009 were included in this retrospective study. The pre- and post-neoadjuvant chemotherapy (NCT) treatment expression levels and changes of Ki-67 proliferation index, estrogen receptor (ER), progesterone receptor (PgR), epidermal growth factor receptor 2 (HER-2), cyclin D1, and nm23-H1 were detected by immunohistochemistry (IHC). The pCR rate was 9.1% (95% CI, 5.3–12.9%). In univariate analysis, poor tumor differentiation, OR after 2 cycles of NCT, both negative of ER/PgR, negative HER-2, positive cyclin D1, and positive nm23-H1 were found to be significantly predictive of a pCR. Histological grade and ER/PgR status were significant for pCR on multivariate analysis (P = 0.023 and 0.003, respectively). The expression levels of cyclin D1 (median, 8% vs. 9%; P = 0.016) after NCT treatment increased significantly, while the median Ki-67 proliferation index was dramatically decreased after NCT treatment from 35 to 15% (P = 0.036). However, after a Bonferroni adjustment, only the difference of Ki-67 proliferation index was still significant (P = 0.026). Histological grade and ER/PgR status are independent predictive factors of pCR to neoadjuvant docetaxel plus epirubicin combination chemotherapy in locally advanced breast cancer. Expression of HER-2, Ki-67, cyclin D1, and nm23-H1 were not predictive for pCR.

     

Identification of a second human nm23 gene, nm23-H2

Reduced RNA and/or protein levels corresponding to the murine nm23-1 and human nm23-H1 complementary DNA clones have been correlated with high tumor metastatic potential in several rodent model systems and human breast carcinomas. We report the identification of a second human nm23 gene, designated nm23-H2. The pNM23-H2S complementary DNA clone predicted a Mr 17,000 protein 88% identical to nm23-H1. nm23-H2 also shared a significant homology with nucleoside diphosphate kinases and a Drosophila developmental gene. Southern blots containing BglII-restricted genomic DNA, which exhibited an allelic restriction fragment length polymorphism for nm23-H1, contained nonallelic bands upon rehybridization to the nm23-H2 probe. Thus, nm23-H1 and nm23-H2 are distinct genes. Northern blot hybridization of nm23-H1- and nm23-H2-specific probes to breast tumors and cell lines indicated that nm23-H1 expression was reduced in high metastatic potential tumor cells to a greater extent than nm23-H2. The data indicate the existence of a family of independently regulated nm23 genes.     

膀胱癌中nm23-H1基因突变及表达的意义

目的：探讨ｎｍ２３－Ｈ１基因在膀胱癌中突变及表达的意义。方法：应用半定量逆转录聚合酶链反应（ＲＴ－ＰＣＲ）和银染单链构象多态性（ＳＳＣＰ）方法检测ｎｍ２３－Ｈ１基因在２５例膀胱癌组织及１５例对照组织中突变和表达情况。结果：对照粘膜中未检测出ｎｍ２３－Ｈ１基因突变,而在２５例膀胱癌组织中发现６例出现ＰＣＲ产物单链泳动状态异常,异常率为２４％。癌组织和对照组织均有ｎｍ２３－Ｈ１基因ｍＲＮＡ的表达,８８     

Immunoreactivity of p53, Ki-67, and c-erbB-2 in phyllodes tumors of the breast in correlation with clinical and morphologic features

BACKGROUND AND OBJECTIVES: Phyllodes tumor (PT) is a biphasic tumor with unpredictable behavior. Our study aimed to evaluate clinicopathologic factors and biomarkers that may be helpful in predicting the outcome of these tumors. METHODS: We evaluated immunoreactivity of p53, c-erbB-2, and Ki-67 in 23 PT treated over a 10-year period. The proliferative activity in PT and expression of p53 and c-erbB-2 were correlated with clinicopathologic features of the tumors and patients' outcome. RESULTS: Positive stromal p53 immunoreactivity was found in PT with atypia, infiltrative borders, high cellularity, as well as in PT that displayed higher then average proliferation index, although none of these parameters reached statistical significance. There was a good correlation between proliferative stromal cell activity expressed Ki-67-labeling index and the malignant features of the tumors. Primary tumors that recurred displayed high proliferative activity. Three of four recurrent tumors showed a progression toward higher malignant phenotype with concomitant increase in proliferative stromal cell activity. c-erbB-2-positive tumors had no particular histologic features or association with either p53 positivity or higher proliferative indices. CONCLUSIONS: p53 expression tends to be more frequent in PT with higher malignant potential but did not predict recurrence. Incompletely excised tumors that recurred displayed high proliferative activity in their primary tumors. Progression toward more malignant phenotype in the recurrent PT was accompanied with increase in stromal cell proliferative activity, suggesting the presence of biological continuity between benign, borderline, and malignant PT.     

Immunohistochemical detection of nm23-H1/NDP kinase in childhood thyroid carcinoma

Childhood thyroid cancer is known to be aggressive. High incidence of lymph node and distant metastasis are characteristic features of these cases. In adult, reduced expression of nm23-H1/nucleoside diphosphate (NDP) kinase has been correlated with cancer invasion and metastasis in some tumor types. Therefore, we examined the expression of nm23-H1 gene product in childhood thyroid carcinomas in Japan. 27 primary thyroid carcinomas and 8 metastatic lymph nodes were analyzed by immunohistochemistry using monoclonal antibody H1-229. 21 out of 23 cases (91%) of papillary carcinomas were positively immunostained, whereas none of the 4 follicular carcinomas showed any immunoreactivity. No correlation was found between the nm23-H1/NDP kinase antigen expression and nodal involvement or distant metastasis in primary tumors. However, only 50% (4 out of 8) of metastatic lymph nodes from papillary carcinoma were positively stained, demonstrating a significant decrease comparing to those of primary sites. These data indicate that the expression of nm23-H1/NDP kinase cannot predict tumor metastatic potential in childhood thyroid cancer.     

Expression of nm23-Hl and nm23-H2 Proteins in Prostate Carcinoma

The nm 23 gene products/nucleoside diphosphate (NDP) kinase expression in prostate carcinomas and benign hyperplasias was evaluated immunohistochemically. Monoclonal antibodies against nm 23-H1 and nm 23-H2 proteins were prepared using the corresponding proteins fused with glutathione S-transferase as immunogens. Of the 80 cases of nonmetastatic prostate carcinoma examined, 74% (59/80) and 60% (48/80) were immunoreactive for nm 23-H1 or nm 23-H2 protein, respectively. Negative staining for nm 23-H1 occurred in 83% of metastatic lesions, while 34% were negative for nm 23-H2. All primary tumors corresponding to the metastases examined showed positive immunostaining for nm 23-H1, indicating an inverse relationship between expression of this protein and metastatic status. nm 23-H2 protein was detected in 83% of primary tumors and its expression appeared to he significantly correlated to the degree of histological differentiation. In contrast, all cases of benign prostatic hyperplasia showed elevated levels of both nm 23-H1 and nm 23-H2 expression. These data suggest that the nm 23/NDP kinase may play a role in suppressing the expression of malignant potential in prostate carcinomas.     

nm23-H1基因对肺癌细胞株恶性表型的逆转作用

目的：通过稳定、高效的基因转染方法将nm23-H1基因导入nm23-H1基因缺失的人肺大细胞癌细胞株L9981,探讨nm23-H1基因是否具有逆转肺癌恶性表型的功能。方法：利用阳离子脂质体介导的基因转染技术,将nm23-H1基因导入L9981细胞株;利用细胞生长曲线、克隆形成率、Boyden小室法等方法研究转染前后细胞体外增殖、黏附、侵袭、转移能力的改变。结果：转染后的L9981细胞株中有nm23-H1基因mRNA和蛋白的阳性表达;转染后细胞株体外增殖速度减慢,克隆形成率降低,黏附性以及侵袭性均降低。结论：nm23-H1基因对肺癌细胞株L9981的体外生长、侵袭、转移具有负调控作用,表明nm23-H1基因具有逆转肺大细胞癌恶性转移表型的能力,为应用nm23-H1基因治疗肺癌提供了客观依据。     

乳腺癌组织p16与nm23-H1的表达及临床病理相关性研究

目的 :探讨乳腺癌组织p16与nm2 3 H1的表达及其与临床病理特征和预后的相关性。方法 :免疫组化SP法检测 12 0例乳腺癌组织P16与nm2 3 H1的表达 ,并结合临床病理特征及预后进行分析。结果 :乳腺癌组织p16阳性率 5 5 0 0 % ,p16阳性表达与腋淋巴结转移负相关 ,P <0 0 5 ,与 5年生存率正相关 ,P <0 0 5 ;nm2 3 H1阳性率 4 2 5 0 % ,nm2 3 H1阳性表达与腋淋巴结转移负相关 ,P <0 0 5 ,与 5年生存率正相关 ,P <0 0 5。p16与nm2 3 H1表达无相关性 ,p16 (- ) nm2 3 H1(- )淋巴结转移率高于p16 (+) nm2 3 H1(+) ,5年生存率低于p16 (+) nm2 3 H1(+) ,统计学检验差异具有显著意义 ,P <0 0 0 1;P <0 0 1。结论 :p16与nm2 3 H1阳性表达与乳腺癌的预后有关 ,其联合表达可更好地判断乳腺癌的转移和预后     

Prognostic significance of p53, nm23, PCNA and c-erbB-2 in gastric cancer

BACKGROUND: Although the TNM stage is the most important prognostic factor for gastric cancer, there is a need for new prognostic and predictive factors, because the prognosis varies among patients of the same stage. The purpose of this study was to clarify the relationship of p53, nm23, proliferating cell nuclear antigen (PCNA) and c-erbB-2 with the clinicopathological parameters and the survival results. METHODS: For 841 patients who had undergone gastrectomy for gastric cancer at Seoul National University Hospital from July 1996 to December 1997, the expression levels of p53, nm23, PCNA and c-erbB-2 in gastric cancer tissues were examined immunohistochemically. Also, the clinicopathological parameters such as gender, age, operation type, TNM stage and size of the tumor, histology and Lauren classification were analyzed retrospectively. RESULTS: There were 568 males and 273 females (2.07:1) with a mean age of 56 years (range:25-82 years). The percentages of positive expression of p53, nm23 and c-erbB-2 were 43, 74 and 17%, respectively; 59% of tumors expressed PCNA index > or =50. p53 expression was associated with age, gender, tumor size, histology, Lauren classification, stage, nm23 expression, PCNA index >or =50 and c-erbB-2 expression. nm23 expression was associated with age, tumor size, Borrmann type, histology, Lauren classification and stage. PCNA index > or =50 was associated with age, gender, tumor size, Borrmann type, histology, Lauren classification and c-erbB-2 expression. c-erbB-2 expression was associated with gender, Borrmann type, histology and Lauren classification. p53 and nm23 were related with poor prognosis in univariate analysis. nm23 was related with poor prognosis of stage III and diffuse-type gastric cancer in univariate subgroup analysis. However, in a multivariate study, these prognostic impacts were not maintained. CONCLUSION: The expression of p53 and nm23 seems to be related with poor prognosis of gastric cancer patients who have undergone gastrectomy. However, the prognostic significance was not revealed by a multivariate analysis.     

Estrogen Receptor Mutations and Changes in Estrogen Receptor and Progesterone Receptor Protein Expression in Metastatic or Recurrent Breast Cancer

To investigate the frequency of estrogen receptor ( ER ) gene mutation in metastatic or recurrent breast cancer, metastatic lymph nodes or recurrent breast cancer tissue from 35 patients with ER-positive primary tumors were screened for mutations in the hormone-binding domain of the ER gene by sequence analysis. Four missense mutations, Val316Ile, Gly344Val, Ala430Val and Gly494Val, were identified in these lesions. Second, to clarify whether there is any disparity in hormone receptor status between primary and metastatic or recurrent tumors, we immunohistochemically studied 117 specimens including the above 35 specimens obtained from metastatic or recurrent breast cancer patients using monoclonal anti-ER and progesterone receptor (PgR) antibodies. Although hormone receptor status, especially ER, was highly maintained through disease progression, negative change in PgR expression at relapse (33%) was identified more frequently than in metastatic lymph nodes (6.7%). Therefore, it was suggested that development of PgR-negative phenotype might correlate with disease progression in some breast cancer patients. These results suggest that ER mutations in metastatic or recurrent breast cancer may be more frequent than in primary lesions, irrespective of high maintenance of ER protein expression through disease progression.     

nm23-H1 protein immunohistochemical expression in human breast cancer

A series of 76 patients undergoing surgery for primary breast carcinoma has been prospectively studied in order to evaluate the relative weight of nm23-H1 protein expression in disease-free survival. Expression of nm23 protein was immunohistochemically assessed. In all, 39% (29/74) of the turners showed positive staining for nm23-H1 protein expression. Negative nm23-H1 expression was found in poorly differentiated, tumors (p<0.02). There was no significant relationship between nm23-H1 and the other clinicopathological and biological features examined. In the univariate statistical analysis, node positivity, G3 histological grade and high flow cytometric S phase fraction (SPF) value proved to be significantly related to risk of relapse. In the multivariate analysis, only histological grade (G3) and high SPF values (>10.6) proved to be independently related to risk of relapse, with a hazard ratio of 9.84 and 7.98 respectively. Our preliminary study suggests that immunohistochemical nm23-H1 expression should not be considered a marker for predicting tumor progression and patient prognosis.     

nm23-H_1mRNA表达与肾母细胞瘤临床行为的关系

应用Northernblot分子杂交技术，检测30例肾母细胞瘤及其对应正常肾组织中nm23－H1mRNA表达。通过nm23－H1mRNA水平的比较及其与临床病理的分析，发现肾母细胞瘤nm23－H1mRNA呈低表达，随着病程的演进和转移的出现，肿瘤细胞中nm23－H1mRNA表达水平呈显著性降低，与肿瘤的病理类型无显著相关性（P＞0．05）。提示nm23－H1RNA低表达可能是肾母细胞瘤演进和转移的重要因素。     

Proliferating cell nuclear antigen (pcna) and C-erbb-2 oncoprotein in breast-carcinoma with correlations to histopathological parameters and prognosis

Overexpression of PCNA (more than 25% positive tumour cells) and positivity of c-erbB-2 oncoprotein were immunohistochemically demonstrated in 490 formalin-fixed and paraffin-embedded breast carcinomas. Overexpression of PCNA and c-erbB-2 correlated with large tumour size, presence of lymph node metastases, high histological grade (poor differentiation), and absence of steriod hormone receptors features indicating an aggressive phenotype. In univariate analysis overexpression of PCNA correlated with poor overall survival (p<0.05), whereas c-erbB-2 was of no prognostic significance. In multivariate analysis both PCNA and c-erbB-2 failed to be of independent prognostic significance. In order to identify women with different prognosis an index termed immunoscore, based upon the results of the immunoreactivity of both PCNA and c-erbB-2 was constructed. The immunoscore was correlated with tumour size, lymph node status, histological grade, and steroid hormone receptor status. In univariate analysis of survival data the immunoscore was a prognostic parameter of poor overall survival. In multivariate analysis the classical histopathological parameters such as tumour size, histological grade and progesterone receptor status turned out to be of independent prognostic significance. The immunoscore was associated with poor prognosis but did not reach independent statistical significance (p=0.08). Further studies including a larger number of patients must be carried out in order to determine the prognostic significance of the immunoscore in multivariate analysis.     

Breast-cancer in the elderly - detection and treatment modalities in 341 women

We examined nm23-H1 protein levels in human oligodendrogliomas by immunohistochemistry. This class of brain tumor does not form spontaneous metastases, but its progression from benign (oligodendroglioma) toward malignant phenotype (oligodendroglioma transitionale and glioblastoma oligodendrogliale) can be followed. Two types of tumors, ODG-II and ODG-T, were highly positive for nm23 protein. However, there was no clear correlation between the extent of protein expression and tumor aggressiveness. No nm23 protein was detected in nonproliferative normal brain tissues and was found in only a few ODG-I specimens. As cell proliferation becomes more pronounced (OGD-II, ODG-T), nm23 protein becomes detectable in almost all samples. However, of the glioblastoma oligodendrogliale samples examined, 76% were negative for nm23-H1 protein. suggesting a change in nm23-H1 gene expression with increasing neoplastic progression. Our findings are in contrast to a proposed role of nm23-H1 protein as a tumor metastasis suppressor and support that it cannot serve as a reliable prognostic tumor indicator in all cases. However, our findings may contribute to a better understanding of glial tumor development and improve the accuracy of tumor diagnosis.